Hi all
Sorry for the late response, but I heartily agree that you should not
crush the material until you return. Just removing your flower from the
plant should start a series of cell death responses that could damage
nucleic acids. Crushing the fresh tissue will release a flood of acids
and waste products that will decrade the nucleic acids, let alone all
the RNases that will degrade your RNA within hours at ambient
temperatures.
I would try RNAlater (from Ambion, but no affiliation) for 1 day at
ambient temp and get the samples into frig or freezer within 1 day. I
don't know if you can get RNA from dried tissue, but if your trip into
the jungle will be for a prolonged period of time without any
refrigeration, you could probably dessicate the tissue samples and get
DNA at a later time. I have also isolated good quality DNA from plants
and insects stored in ETOH. I think you may need to consider working
with DNA.
Hope this helps
Deanne Bell
USDA Agricultural Research Service
Crop Diseases, Pests & Genetics
9611 South Riverbend Avenue
Parlier, CA 93648
voice (559) 596-2806
fax (559) 596-2897
dbell at fresno.ars.usda.gov
>-----Original Message-----
>From: methods-bounces at oat.bio.indiana.edu>[mailto:methods-bounces at oat.bio.indiana.edu] On Behalf Of Bo Johansen
>Sent: Tuesday, August 16, 2005 1:00 AM
>To: methods at magpie.bio.indiana.edu>Subject: Protecting RNA in the jungle
>>Hi
>>I am going on a trip to collect RNA samples from flower buds
>in the jungle of plants that cannot be cultivated. It is
>impossible to bring liquid nitrogen on the trip, and I won't
>get into a lab for 2-3 weeks.
>What do I do.
>>I have thought of collecting the material in Eppendorph tubes,
>add 1 ml og 2.5 M LiCl and crush the material with a pistil.
>Close the tube and leave it until I get to a lab
>>Or
>>Collecting the material in an Eppendorph tube, add 1 ml og 4M
>guanidine isothiocyanate, 25mM sodium citrate, pH7, 0.5%
>sarcosyl, 100mM b-mercaptoethanolm, crush the material and
>leave it until I get to a lab.
>>Will any of these methods leave the RNA intact or are there
>other methods?
>>Bo
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